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Biomedical subjects

J A McIntyre

Publications and source records attributed to J A McIntyre.

At least 73 records · Page 4Linked to original sources
PubMed1993

Predominant expression of the beta subunit of prolyl 4-hydroxylase (disulfide isomerase) in human extravillous trophoblasts.

Prolyl 4-hydroxylase is a heterodimeric enzyme that is crucial in the biosynthesis of collagen. The beta subunit of this enzyme is a multifunctional protein which is also known as protein-disulfide isomerase. Immunofluorescence and monoclonal antibody (Mab) 5B5 were used to localize the beta subunit in human extraembryonic tissues. The strongest sites of 5B5 reactivity were extravillous cytotrophoblasts in the basal plate, uteroplacental arteries and amniochorion, syncytiotrophoblast displayed variable weaker reactivity. Only a small fraction of placental 5B5 antigen was detected as a component of prolyl-4-hydroxylase by affinity chromatography on immobilized polyproline. The results indicate a difference in the expression of an endoplasmic reticulum marker between villous and extravillous trophoblast. The predominance of 5B5 antigen in extravillous trophoblast could be associated with an increased ability to synthesize collagen or other enzymatic reactions associated with prolyl 4-hydroxylase beta subunit.

Antibody Specificity↗
PubMed1993

Immunological studies of lactoferrin in human placentae.

Lactoferrin (LF) and transferrin (Trf) are glycoproteins with strong affinities for ferric ions. Human syncytiotrophoblastic membranes analyzed by enzyme linked immunosorbent assay (ELISA) and immunoblotting were negative with monoclonal and polyclonal antibodies to LF. Immunohistological studies of 35 normal placentae showed that LF was absent from the trophoblast basement membranes, stroma and fetal stem vessel endothelium, but positive cells were occasionally noted in intervillous spaces and fetal stem vessels. In contrast, many LF-positive cells were identified within areas of immunopathology identified by the presence of T cells, HLA-DR-positive macrophages and platelets. Double-antibody experiments showed that the LF-positive cells in these areas reacted with CD15 and CD16 monoclonal antibodies (mAbs), indicating that the cells were polymorphonuclear neutrophils (PMN). PMN from peripheral blood analyzed by flow cytometry and immunocytology also showed reactivities with anti-LF, CD15 and CD16 and we consistently found that circulating PMN reacted better than placental PMN with antibodies to MHC class I antigens and gp 100, (CD67), which is a neutrophil activation marker. PMN adherent within placentae had no detectable MHC class I or CD67 antigens. These findings suggest PMN adherent to placental tissues down-regulate or alter plasma membrane markers. LF appears to play a role in placental inflammation, for LF-positive cells were significantly enriched in areas of immunopathology.

Antibodies, Monoclonal↗
PubMed1993

Phenotypic characterization of normal human placental mononuclear cells.

The placenta is a rich source of immunocompetent cells. We have studied the phenotype, number and origin of placental mononuclear blood cells isolated from 32 normal term placentae using 4 color flow cytometry. Respective maternal and cord blood leucocyte preparations were also compared. Placental tissue without extraembryonic membranes was cut into small pieces and divided. One portion was washed extensively with ice-cold PBS. Both tissue portions were disrupted in a blender and cells were dissociated by using a 180 mu sieve. Leucocytes were isolated by Ficoll-Hypaque density gradient centrifugation. Maternal and cord bloods were HLA typed and in cases of HLA-A2 or B7/40 disparity, monoclonal anti-HLA antibodies to these antigens showed that unwashed placental tissue contained 35% maternal and 65% fetal cells. This ratio, however, was not reflected for a given cell phenotype. In comparison, washed placental tissue contained cells of fetal origin only. Both unwashed and washed placental tissue contained fewer CD3 and CD4, but more CD8 cells than maternal and cord blood. Markers of NK cells such as, CD16, CD56, and CD57 showed this cellular phenotype to be 15 times more abundant in the placental preparations than in cord and maternal blood. The quantitative differences between peripheral blood and placental CD8 and NK cells were further explored with an antiprogesterone receptor antibody in combination with anti-CD8, anti-CD57 and anti-HLA-DR. The number of progesterone receptor (PGR) positive cells was three times higher in placental tissues than in cord or maternal blood. These data indicate that the phenotypic frequencies of certain placental leucocytes are significantly different from maternal and fetal peripheral blood. Progesterone and the presence of PGR may be important in the differential retention of placental leucocytes.

Antigens, CD↗
PubMed1993

Heparin and pregnancy in women with a history of repeated miscarriages.

Many patients who experience recurrent spontaneous abortions (RSA) have high titered antibodies to trophoblast antigens and/or negatively charged phospholipids. Pregnancy success has been documented in some of these patients subsequent to heparin treatment. The effect of heparin on in vitro assays used to detect antibodies to phospholipids and trophoblast antigens has been investigated. We have found that complement-mediated detection of antibodies to trophoblast and lymphocyte antigens is inhibited by absorption of sera with solid-phase heparin. The data show that such inhibition involves an inhibitor and a heparin-sensitive regulator of the inhibitor. Heparin inhibits binding of antiphospholipid antibodies to phospholipids in solid-phase ELISA. The mechanism for this inhibition appears to involve an interaction between heparin and antibody to phospholipid. These findings reveal novel heparin-mediated reactions that bridge immunological and hematological interactions. The extent of heparin modulation of these antibody reactivities should be clarified in future clinical trials involving heparin treatment for RSA.

Abortion, Habitual↗
PubMed1993

Expression of CD59, a human complement system regulatory protein, in extraembryonic membranes.

CD59 (leukocyte cluster of differentiation antigen 59), is a phosphatidylinositol glycan-anchored membrane protein that inhibits lysis of cells by terminal complement system components. To further define complement regulatory proteins relevant to pregnancy, this study characterized the expression of CD59 in human extraembryonic membranes. Immunohistology with CD59 monoclonal antibody MEM-43 showed that this molecule was normally present on the apical surface of the syncytiotrophoblast, on extravillous cytotrophoblast, and amniotic epithelium. Immunoblotting confirmed that first and second trimester syncytiotrophoblast microvilli (STM) contained a glycoprotein similar in mass and glycosylation to CD59 from adult cells and tissues. Reactivities of STM with MEM-43 in ELISA were 2- to 6-fold higher than those of kidney, erythrocyte and platelet membranes. Term placental STM from recurrent spontaneous aborting patients after immunotherapy, reacted with MEM-43 in ELISA similarly to STM from normal individuals. Plasmas from pregnant women and umbilical cords had 50% or greater reactivity with MEM-43 than did normal plasmas. CD59 could help protect extraembryonic epithelia from damage by complement in maternal blood and amniotic fluid. The apical location of CD59 reflects the immunological roles and functional polarization of plasma membranes in the syncytiotrophoblast.

Abortion, Spontaneous↗
PubMed1993

Nitric oxide release in response to gut injury.

We evaluated nitric oxide release in several models of intestinal inflammation through luminal nitrite concentrations. In anesthetized rabbits, piglets, and guinea pigs, luminal lavages were collected from loops of normal or injured small intestine. Lavages were analyzed spectrophotometrically for nitrite (Griess reagent) and protein. Myeloperoxidase (MPO) content of intestinal segments was used as an index of granulocyte infiltration and intestinal inflammation. Acute ileal inflammation was induced by luminal acetic acid + casein in rabbits and luminal deoxycholate in neonatal piglets and adult rabbits. Chronic ileitis was induced in guinea pigs by intraluminal trinitrobenzenesulfonic acid. In each model nitrite levels in ileal lavages were significantly greater than control loops/animals. Increased luminal protein and intestinal MPO activity paralleled the changes in nitrite levels. To determine whether nitric oxide production influenced mucosal repair, segments of ileum were perfused with the L-arginine antagonist NG-nitro-L-arginine methyl ester (L-NAME, 10 mg/ml) after acute acetic acid + casein exposure. L-NAME administration reversed the decline in epithelial permeability characteristic of epithelial restitution, causing an increase in epithelial permeability which was readily reversible. These results suggest that nitrite production is a useful index of gut injury and that nitric oxide may contribute to the functional repair of the epithelial barrier under acute conditions.

Animals↗
PubMed1993

Integrating psychology and obstetrics for medical students: shared labour ward teaching.

Two studies relating to the inclusion of psycho-social issues in the training of obstetricians are reported here. The first reports on the extent to which currently practising obstetricians have received training in these aspects. The second explored the value of an innovative teaching approach combining psychological and obstetric training for medical students in the labour ward. A postal survey with responses from 220 obstetricians and paediatricians revealed that little information on psychological aspects of obstetric practice had been included in their undergraduate or postgraduate training or obtained from voluntary continuing education programmes. Experience was the primary source of training in these subjects. The second study explored the impact of joint psychological and obstetric teaching ward rounds for medical students. Students attending these integrated sessions reported being better prepared for the psycho-social aspects of obstetrics and showed a greater awareness of cross-cultural differences in needs of women during birth.

Cross-Cultural Comparison↗
PubMed1993

Guinea pig ileitis is attenuated by the leumedin N-(fluorenyl-9- methoxycarbonyl)-leucine (NPC 15199).

Anti-inflammatory properties have been ascribed to a series of N-(fluorenyl-9-methoxycarbonyl) amino acids called leumedins that inhibit the activity of granulocytes and T-lymphocytes. We evaluated one of these leumedins, N-(fluorenyl-9-methoxycarbonyl) leucine (NPC 15199), in a model of ileitis in guinea pigs. Ileitis was induced by intraluminal trinitrobenzenesulfonic acid (TNBS 30 mg/kg in 50% ethanol) in anesthetized guinea pigs. NPC 15199 was administered daily (10 or 100 mg/kg, s.c.). After 7 days, the guinea pigs were anesthetized, and saline was administered intraluminally into an ileal loop created at the site of TNBS administration and was withdrawn after 30 min. The changes in lavage protein, nitrite levels, myeloperoxidase (MPO) activity and mast cell numbers were used as indices of inflammation and injury. NPC 15199 (10 or 100 mg/kg) attenuated or abolished TNBS-induced elevations in lavage protein and nitrite content. Only the high dose of NPC 15199 (100 mg/kg) attenuated ileal MPO activity and mast cell hyperplasia. Histological disturbances induced by TNBS administration included crypt hypertrophy, mucosal and submucosal fibrosis and smooth-muscle hyperplasia. These disturbances were reversed by high-dose NPC 15199 (100 mg/kg) but were minimally affected by low-dose NPC 15199 (10 mg/kg). We conclude that NPC 15199 prevents mucosal injury and dysfunction in this model of intestinal inflammation. Inhibition of granulocyte infiltration does not appear to be essential for the beneficial effects of NPC 15199 and suggests that the alternative actions of NPC 15199 may be pertinent to this model.

Animals↗
PubMed1992

Glycosylation of membrane cofactor protein (CD46) in human trophoblast, kidney and platelets.

Many cell surface glycoconjugates are differentiation markers and are involved in cell-cell and intermolecular interactions in development, immunity and cancer. Membrane cofactor protein (MCP) comprises structurally related 65 and 55 kDa glycoproteins that bear O- and N-linked glycans. MCP prevents amplification of autologous complement action on human cells. We used immunoblotting with MCP-specific monoclonal antibody TRA-2-10 to determine lectin-binding properties and glycosidase sensitivities of MCP in a study of cell-specific variation in glycosylation of this protein. The results showed that N-linked glycans on placental syncytiotrophoblast and cytotrophoblast, kidney and platelet MCP are similar in binding to concanavalin A and Lens culinaris lectins, but are not bound by leucophytohemagglutinin. Lectin binding prior to and after neuraminidase digestion indicates that MCP from these sources is highly sialylated. 65 kDa MCP was confirmed to contain more O-linked glycans than 55 kDa MCP. A fraction of platelet 65 kDa MCP is distinct, however, in bearing peripheral fucose residues. Syncytiotrophoblast is unique in containing a 110 kDa form of MCP in non-reducing SDS-PAGE that resembles 65 kDa MCP in glycosylation. Chorion laeve MCP in 4 of 8 preparations was unusually heterogeneous and differed from syncytiotrophoblast MCP after neuraminidase digestion in the forms bound to peanut agglutinin and WGA. The results indicated for the first time, differences in O-linked glycosylation of MCP in chorion laeve cytotrophoblast relative to syncytiotrophoblast, platelet and kidney MCP. We conclude that structures of MCP glycans can differ between trophoblasts and other cell types.

Antigens, CD↗
PubMed1992

Previous birth outcome of antenatal clinic attenders in northern KwaZulu--perinatal and infant mortality rates.

A questionnaire survey was undertaken among pregnant women presenting for the first time to antenatal clinics attached to Mosvold Hospital in rural northern KwaZulu. They were asked details of the outcome of their previous pregnancies and the survival of their last-born children. Of 2,388 mothers interviewed, 1,795 had given birth previously. Of these, 41% had had their last deliveries at home, 47% in hospital and 10% at clinics. The perinatal mortality rate was 38/1,000, with no significant differences between home and hospital or clinic births. The infant mortality rate for live-born children was 62/1,000. Fifty per cent of child deaths occurred at home. The study methodology was easily applied and provided an acceptable alternative technique for measuring child mortality rates, which are so difficult to obtain in rural areas in southern Africa.

Ambulatory Care Facilities↗
PubMed1992

AIDS--the Baragwanath experience. Part I. Epidemiology of HIV infection at Baragwanath Hospital, 1988-1990.

This four-part series describes the experience with HIV infection at Baragwanath Hospital to December 1990. In this first part we give an overview of the emergence of this disease and of its impact on the hospital. From July 1988 to December 1990, 426 HIV-positive individuals were identified: 58 were women identified in surveys at antenatal and gynaecology clinics, 60 were parents of infected babies or sexual contacts of hospitalised patients, and 30 were not inpatients (mainly patients at Soweto clinics). Of the 278 inpatients, 16 were identified in the latter 6 months of 1988, 54 in 1989 and 208 in 1990. Fifty-one per cent of the patients were in the adult medical wards, 11% in surgery, 20% in paediatrics and 18% in other disciplines. One per cent of sera from Wassermann reaction-positive women in mid-1990 were HIV-positive; this also applied to 1% of antenatal women in late 1990 and 2.7% of women with pelvic inflammatory disease. The HIV antigen assay is a useful adjunctive assay in the evaluation of HIV-antibody-positive children. The false/true-positive ratio of a rapid HIV antibody test was 2:1 initially, but the specificity improved with experience. The use of rapid diagnostic assays for HIV must be restricted to laboratories with experience in reading assays and where definitive follow-up testing is assured.

Acquired Immunodeficiency Syndrome↗
PubMed1992

AIDS--the Baragwanath experience. Part II. HIV infection in pregnancy and childhood.

By the end of 1990, 51 symptomatic children with perinatally acquired HIV infection had been admitted to Baragwanath Hospital. Of 42 children who were followed up for at least 3 months, 15 (35.7%) died and 16 (38.1%) were lost to follow-up. The case fatality rate for these children lies between 35.7% and 73.8%. Most children became symptomatic before the age of 6 months and presenting features seen in over 70% of cases included lymphadenopathy, failure to thrive and hepatomegaly. Surviving children had recurrent admissions to hospital, predominantly for respiratory distress. Many had bacterial pneumonias. Cardiac involvement (cardiomyopathy and cor pulmonale), recurrent serious bacterial infections and neurodevelopmental abnormalities were common. Our experience confirms that vertically acquired HIV infection has a relatively short incubation period and progresses rapidly with cardiorespiratory symptoms predominating. Five HIV seroprevalence studies have been conducted in pregnant women attending Baragwanath Hospital and the Soweto clinics since 1988. The calculated doubling time is between 7 and 21 months. At the end of 1990 the HIV seroprevalence rate in pregnant women was 0.82% (95% confidence limits 0.44-1.19%).

Adolescent↗
PubMed1992

AIDS--the Baragwanath experience. Part IV. Counselling and ethical issues.

This fourth part of the series describes the setting up of counselling for HIV-positive patients in response to the need at Baragwanath Hospital. Nurse counsellors were trained and supervised. The work, extending over 2 1/2 years, raised a number of issues for patient care and also for wider community education. Brief reference is made to pertinent ethical and cultural issues.

Counseling↗
PubMed1992

South African obstetricians' views on caesarean section.

The widespread increase in the rate of caesarean sections performed in the Western world has become a cause for concern. In this country, two systems of health care exist, one provided by the State and the other based on private practice. A high incidence of caesarean sections has been reported, especially among private practitioners. This study investigated doctors' perceptions of caesarean section practices and explored the availability of facilities which could help to reduce the high caesarean section rate. Questionnaires were mailed to all registered obstetricians. Of these 45.2% (233) responded. Findings indicate that private obstetricians are more likely to perform caesarean sections even though they do have good facilities available to monitor or assess difficult deliveries. Other factors influencing their practice are also explored.

Attitude of Health Personnel↗
PubMed1992

Inhibition of immunoglobulin (Ig)G-Fc-mediated cytotoxicity by seminal plasma IgG-Fc receptor III antigens.

OBJECTIVE: To determine if the presence of soluble immunoglobulin (Ig)G-Fc receptor III (Fc gamma RIII) antigens in human seminal plasma interfere with IgG-Fc-mediated effector functions. DESIGN: An antibody-dependent cellular cytotoxicity (ADCC) assay was used as a model for IgG-Fc-mediated effector functions. Human red blood cells (RBC), labeled with 51Cr were sensitized with rabbit anti-RBC and used as targets for peripheral blood leukocyte (PBL) effector cells. Cytotoxicity was measured by assessing the release of 51Cr from RBC. INTERVENTIONS: (1) Seminal plasma was added at different concentrations to the ADCC, and inhibitory effects were measured. (2) The level of seminal plasma interaction in ADCC was studied by comparing ADCC results in the presence and absence of seminal plasma with findings of target and effector cells that had been preincubated with seminal plasma. (3) The role of seminal plasma Fc gamma RIII in inhibiting ADCC was studied by coincubating seminal plasma with monoclonal antibodies (MAs) Leu 11b that block Fc gamma RIII binding sites. Two isotype-matched MA controls were used at identical concentrations. RESULTS: (1) Seminal plasma dose dependently inhibits ADCC. (2) Seminal plasma inhibition of ADCC occurs at the level of IgG-Fc interaction with effector cell Fc gamma receptors. (3) Inhibitory effects of seminal plasma on ADCC can be specifically blocked by coincubating seminal plasma with MAs Leu 11b that block Fc gamma RIII binding sites. CONCLUSIONS: Seminal plasma Fc gamma RIII antigens interfere with IgG-Fc-mediated effector functions. This mechanism could play a beneficial role in controlling potentially harmful antipaternal immune responses.

Antibody-Dependent Cell Cytotoxicity↗
PubMed1991

A vitronectin-receptor-related molecule in human placental brush border membranes.

The heterodimeric vitronectin receptor (VNR) and platelet glycoprotein IIb/IIIa (GPIIb/IIIa) are two members of the integrin family of cell adhesion receptors that share the same beta subunit (GPIIIa). These proteins are involved in binding to vitronectin, fibrinogen and fibronectin and in cytoskeleton-membrane interactions. The present study shows that the human placental syncytiotrophoblast brush border membrane contains a heterodimer of subunit Mr values of 140,000 and 90,000 (non-reduced) or 125,000 and 100,000 (reduced). This protein was recognized by a monoclonal antibody to GPIIIa, rabbit antisera to the VNR and a human alloantiserum to GPIIIa. Brush border VNR-related protein bound to an immobilized peptide containing the Arg-Gly-Asp sequence and, less avidly, to immobilized fibrinogen. Only a small fraction of brush border VNR was associated with a cytoskeleton fraction. Membrane-bound brush border GPIIIa was distinct from that of platelets in its resistance to digestion by trypsin and Staphylococcus aureus V8 protease, and had a slightly lower mobility on SDS/PAGE. In addition, lectin-binding studies indicate glycosylation differences between microvillar and platelet GPIIIa heterodimers. Thus, although placental syncytiotrophoblast expresses a beta 3 integrin in its apical brush border, differences in protease sensitivity and carbohydrate content suggest that it may lack or mask certain antigenic determinants. This may be beneficial in avoiding harmful maternal alloantibody responses during pregnancy. Immunohistology showed that the VNR was present in syncytiotrophoblast apical but not basal plasma membranes, and was absent from other forms of trophoblast. The brush border VNR could function in localizing Arg-Gly-Asp-sequence-containing plasma proteins to the materno-trophoblastic interface.

Blood Platelets↗